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Product Details:
Payment & Shipping Terms:
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Shelf Life: | 12 Months | Sample Performance: | Tissue, Serum, Honey, Egg |
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Specifications: | 96 Wells/kit | Incubation Time: | 30min-15min |
Sensitivity: | 1ppb | Incubation Temperature: | 25℃ |
MOQ: | 1 Kit | ||
High Light: | Fluoroquinalone ELISA Kit,honey test kit,Food safety test kit |
Fluoroquinolone(FQNS) ELISA Kit
Catalog No. LSY-10016
1. Principle
This test kit is based on the competitive enzyme immunoassay for the detection of Fluoroquinolone in samples. The coupling antigens are pre-coated on the micro-well stripes. The Fluoroquinolone in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti- Fluoroquinolone antibodies. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the sample has a negative correlation with the Fluoroquinolone in the sample. This value is compared to the standard curve and the Fluoroquinolone concentration is subsequently obtained.
2. Technical specifications
Sensitivity: 1 ppb
Incubation Temperature: 25℃
Incubation Time: 30min—15min
Detection limit
Enrofloxacin (ENR) 1ppb
Ciprofloxacin (CIF) about 1ppb
Ofloxacin (OFL) about 1ppb
Danofloxacin (DAN) about 1ppb
Norfloxacin (NOR) about 0.5ppb
Lomefloxacin (LOM) about 1ppb
Pefloxacin (PEF) about 0.5ppb
Enoxacin (ENO) about 1ppb
Aosuo Li acid (OXO) about 1ppb
Fluoroquinolone acid (FLU) about 1ppb
Ma Paul Sand Star (MAR) about 1ppb
Ammonia difloxacin (AMI) about 1ppb
That difloxacin (NAD) about 2ppb
Cross-reaction rate:
Enrofloxacin (ENR) 100%
Ciprofloxacin (CIF) 92.88%
Ofloxacin (OFL) 98.86%
Aosuo Li acid (OXO) 116.86%
Danofloxacin (DAN) 102.63%
Norfloxacin (NOR) 148.65%
Lomefloxacin (LOM) 86.24%
Pefloxacin (PEF) 156.60%
Enoxacin (ENO) 98.97%
Fluoroquinolone acid (FLU) 96.73%
Ma Paul Sand Star (MAR) 110.63%
Ammonia difloxacin (AMI) 98.86%
That difloxacin (NAD) 56.81%
Recovery rate
Tissue,egg, serum 80±15%
Honey 75±15%
3. Components
1 | Micro-well strips |
12 strips with 8 removable wells each |
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2 | 6× standard solution (1mL each) | 0ppb | 1ppb |
3ppb | 9ppb | ||
27ppb | 81ppb | ||
3 | Enzyme conjugate | 7ml | red cap |
4 | Antibody working solution | 7ml | blue cap |
5 | Substrate A solution | 7ml | white cap |
6 | Substrate B solution | 7ml | black cap |
7 | Stop solution | 7ml | yellow cap |
8 | 20× concentrated washing buffer | 40ml | white cap |
9 | 2× concentrated redissolving solution | 50ml | transparent cap |
4. Materials required but not provided
5. Sample pre-treatment
Instructions (The following points must be dealt with before the pre-treatment)
1) This test kit can detect tissue sample: animal tissue, poultry, aquatic. Eg: Chicken, duck, bovine, rabbit, fish, shrimp etc.
Solution preparation before sample pre-treatment:
Vacetonitrile-Vmethylene chloride = 1 : 4
100ml Acetonitrile (CH3CN) -Methylene chloride mixing solution (Vacetonitrile-Vmethylene chloride = 4 : 1), add 5ml 0.1 M HCl solution.
5.1 Tissues (chicken/liver, pork/liver, fish, shrimp etc.), egg
1) Weigh 2.0 ± 0.05 g of the homogenized tissue sample into 50 ml centrifuge tube
2) Add 8 ml of the Acetonitrile (CH3CN) -Methylene chloride mixing solution, shake for 5 min, centrifuge at above 4000 r/min at 15 ℃ for 10 min
3) Take 4 ml the clear organic phase (upper layer) into a dry tube, blow to dry with nitrogen or air completely by rotary evaporation at 56 ℃
Fold of dilution of the sample: 1
5.2 Serum
1) Use centrifuge tube with heparin sodium (20-30 unit/ml blood) to collect chicken blood sample (Suggestion: blood collection syringes are recommended rinsing with heparin).Place the blood sample in the room temperature for 1 hour. After obtain plasma, centrifuge at above 4000r/min at 15 ℃ for 10 min, take out 1 ml plasma.
2) Add CH3CN (without water) 4 ml , mix up-and-down thoroughly for 5 min, centrifuge at above 4000r/min at 15 ℃ for 10 min.
3) Move the clear supernatant (upper layer) to another centrifuge tube, add 2ml 0.02M PB buffer, mix evenly.
4) Add 5 ml Methylene chloride, mix evenly for 5 min, centrifuge at above 4000r/min at
15 ℃ for 10 min, remove the upper layer, take the lower organic phase to dry bottle(clear without impurities), blow to dry with nitrogen or air completely by rotary evaporation at 50 ℃
5) Dissolve the dry residues in 1 mL of the diluted redissolving solution, add 1 mL N-hexane, mix for 30 seconds; centrifuge at above 4000 r/min at 15℃ for 5 min.
6) Absorb out lightly the upper and middle layer white impurities, take lower phase 100µl, add 100µl diluted redissolving solution, mix for 30s.
7) Take 50µl solution for further analysis.
Fold of dilution of the sample: 2
5.3 Honey
Fold of dilution of the sample: 2
6. ELISA procedures
6.1 Instructions
6.2 Operation procedures
7. Result judgment
There are two methods to judge the results; the first one is the rough judgment, while the second is the quantitative determination. Note that the OD value of the sample has a negative correlation with the content of Fluoroquinolone in the sample.
7.1 Qualitative determination
The concentration range (ng/mL) obtained from the comparison the average OD value of the sample with that of the standard solution. Assuming that the OD value of the sampleⅠ is 0.238, and that of the sampleⅡ is 0.946, the OD value of standard solutions is: 1.845 for 0 ppb, 1.542 for 1 ppb, 1.130 for 3 ppb, 0.635 for 9 ppb, 0.326 for 27 ppb ,0.156 for 81 ppb, accordingly the concentration range of the sampleⅠ is 27 to 81 ppb, and that of the sampleⅡ is 3 to 9 ppb. (multiplied by the corresponding dilution fold)
7.2 Quantitative determination
The mean values of the absorbance values is equivalent to the percentage of the average OD value (B) of the sample and the standard solution divided by the OD value (B0) of the first standard solution (0 standard) and subsequently multiplied by 100%, that is,
Percentage of absorbance value = | B | ×100% |
B0 |
B—the average (double wells) OD value of the sample or the standard solution
B0—the average OD value of the 0ng/mL standard solution
Draw the standard curve with the absorption percentages of the standard solutions and the semilogarithm values of the Fluoroquinolone standard solutions (ng/mL) as Y- and X-axis, respectively. Read the corresponding concentration of the sample from the standard curve by incorporating its absorption percentage into the standard curve. The resulting value is subsequently multiplied by the corresponding dilution fold, finally obtaining the Fluoroquinolone concentration in the sample.
Using the professional analyzing software of this kit will be more convenient for the accurate and rapid analysis of a large amount of samples. (Please contact us for this software)
8. Precautions
8. The optimum reaction temperature is 25 ℃, and too high or too low temperatures will result in the changes in the detecting sensitivity and OD values.
9. Storage and expiry date
Storage: store at 2-8 ℃, not frozen.
Expiry date: 1 year; date of production is on box.
Note: If the Vacuum package of microplate has leakage, it is still valid to use, do not affect the test result, be relax to use.